New England Biolabs, M0569S, Immobilized T4 DNA Ligase

Looking for additional T4 DNA Ligase formulations and variants? Related Categories DNA Ligases Applications Non-Cloning Ligation,, DNA Ligation,, Cloning Ligation Specification Reaction Conditions 1X T4 DNA Ligase Reaction Buffer 1 T4 DNA Ligase Reaction Buffer 50 mM Tris-HCl 10 mM MgCl2 1 mM ATP 10 mM DTT (pH 7.5 @ 25°C) Usage Concentration 10 mg/ml Storage Buffer 10 mM Tris-HCl 1 mM DTT 0.1 mM EDTA 50 mM KCl 50% Glycerol pH 7.4 @ -20°C Heat Inactivation 65°C for 10 minutes Unit Assay Conditions Product supplied at 10 mg/ml which provides 60 cohesive end units (CEU) per microliter of slurry. FAQ Q: How should Immobilized T4 DNA Ligase (NEB #M0569) be stored and handled? A: Immobilized T4 DNA Ligase should be stored at -20°C. Beads should not be stored at -80°C to avoid multiple freeze/thaw cycles. Upon receipt of the material, we recommend briefly spinning the tube using a mini-centrifuge in order to collect the beads and storage buffer together in the bottom of the tube. Inadvertent mixing during shipment can result in partitioning of beads and buffer. This can lead to the beads drying out in the tube during storage and result in a loss of enzyme activity. The storage buffer contains glycerol for optimal stability, as such, the solution does not “hard freeze” at -20°C, unlike other beads. Also, the beads do not permanently agglomerate or degrade when frozen. Q: How should I mix or resuspend Immobilized T4 DNA Ligase (NEB #M0569) beads prior to use? A: Immobilized T4 DNA Ligase should be removed from -20°C and placed directly on ice before dispensing. Prior to any dispensing event, we recommend briefly spinning the tube in a mini-centrifuge, followed by mixing with a pipette to fully resuspend the beads and achieve homogeneity. Care must be taken as the solution is viscous: submerge only the tip of pipette when aspirating and do not create foam when pipette mixing. Pipette mixing can be performed by setting the volume of your pipette to ~50% of the volume in the tube (to avoid drawing air) and pipetting up and down at least 10 times. Once you have dispensed the intended amount, briefly spin down the tube in a mini centrifuge before returning the product to -20°C for storage.